China ; epidemiology ; Enterovirus A, Human ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; drug therapy ; epidemiology ; etiology ; Humans

OBJECTIVES: To investigate the positive rate of enterovirus (EV) nucleic acid in throat swabs of term late neonates hospitalized during the coronavirus disease 2019 (COVID-19) epidemic and the clinical characteristics of the neonates. METHODS: A single-center cross-sectional study was performed on 611 term late infants who were hospitalized in the neonatal center from October 2020 to September 2021. Throat swabs were collected on admission for coxsackie A16 virus/EV71/EV universal nucleic acid testing. According to the results of EV nucleic acid test, the infants were divided into a positive EV nucleic acid group (8 infants) and a negative EV nucleic acid group (603 infants). Clinical features were compared between the two groups. RESULTS: Among the 611 neonates, 8 tested positive for EV nucleic acid, with a positive rate of 13.1‰, among whom 7 were admitted from May to October. There was a significant difference in the proportion of infants contacting family members with respiratory infection symptoms before disease onset between the positive and negative EV nucleic acid groups (75.0% vs 10.9%, P<0.001). There were no significant differences between the two groups in demographic data, clinical symptoms, and laboratory test results (P>0.05). CONCLUSIONS There is a certain proportion of term late infants testing positive for EV nucleic acid in throat swabs during the COVID-19 epidemic, but the proportion is low. The clinical manifestations and laboratory test results of these infants are non-specific. Transmission among family members might be an important cause of neonatal EV infection.
Infant ; Infant, Newborn ; Humans ; Enterovirus ; COVID-19/diagnosis* ; Cross-Sectional Studies ; Pharynx ; Nucleic Acids ; Enterovirus Infections

Animals ; Critical Illness ; Enterovirus A, Human ; pathogenicity ; Enterovirus Infections ; diagnosis ; therapy ; Hand, Foot and Mouth Disease ; diagnosis ; therapy ; virology ; Humans

In this study, the feasibility of identification and genotypic differentiation of enteroviruses was investigated by using nested reverse transcription-polymerase chain reaction (nested RT-PCR), single-stranded conformation polymorphism (SSCP), and restriction fragment length polymorphism (RFLP) techniques. Two hundred seventy-four clinical samples were assayed by both nested RT-PCR and tube culture method using MRC-5 and MK cells; 58 (86.6%) out of 67 enterovirus culture-positive samples contained enteroviral RNA. In addition, 114 (55.1%) of 207 samples from patients with suspected enteroviral CNS disease with negative viral cultures were positive by the nested RT-PCR. The nested RT-PCR products were genotyped by the SSCP method and the results were compared with serotypes. We could differentiate 6 subtypes, 3 of which are similar to coxsackievirus B3, B5, echovirus 11, plus 3 other subtypes. RFLP cleaved with Sty I, Bgl I, and Xmn I yielded characteristic patterns for each laboratory strains. This study demonstrates the usefulness of the RT-PCR for the rapid diagnosis of enterovirus infection and the potentials of the SSCP method for differentiation of enterovirus strains.
Central Nervous System Diseases ; Diagnosis ; Enterovirus B, Human ; Enterovirus Infections ; Enterovirus* ; Genotype* ; Humans ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; RNA

OBJECTIVETo facilitate early recognition of ominous clinical manifestations, to understand pathophysiology and assess treatment effects in patients with severe enterovirus 71 (EV71) associated hand, foot and mouth disease (HFMD).

METHODA retrospective analysis was performed based on the clinical records, laboratory data and treatment effects which were collected from twelve severe EV71 infected cases from nine hospitals in 2008, in Hubei province, China.

RESULTOf the 12 severe cases, ten (83.3%) were male and two female. The median age was 1.96 yrs (8 m to 7 yrs). The mean hyperthermic duration was 6 days with the peak temperature over 38.5 degrees C, and mean rash duration was 7 days. Fever and rash emerged simultaneously in 4 of 5 cases with cardiopulmonary failure. The severe complications included encephalitis (10 cases), pulmonary edema or hemorrhage (5 cases). Eleven cases were checked with magnetic resonance imaging (MRI) and four cases showed characteristics of encephalitis or meningitis, two with images of naso sinusitis and ethmoid-mastoid inflammation. Chest X-ray examination showed with pulmonary edema on single or both sides (5 cases), bronchitis (4 cases), and normal image (3 cases). There was no specific finding in the cardiac ultrasound and electrocardiogram in any of the patients, as well as the white blood cell count, blood glucose, prothrombin time, partial thromboplastin time and D-dimer. Cerebrospinal fluid showed aseptic meningitis with the increase of cell count in 7 cases. All patients were treated with antibiotics and/or antivirals, such as cephalosporins, ribavirin etc. Eleven patients were treated with intravenous immunoglobulin (total dose 2 - 4.5 g/kg) for 2 - 5 days, and the highest blood concentration of immunoglobulin was detected increasing at 7 g/L. Seven cases were also treated with methylprednisolone 10 - 30 mg/(kg x d), four with dopamine, dobutamine, or digitalis. In addition, by using continuous positive airway pressure by nasal catheter and maintenance of circulation in the cases with cardiopulmonary failure could not relieve the symptoms of dyspnoea, and mechanical ventilation was required to maintain for a mean of 72 hrs (24 - 96 hrs). Except one case died of pulmonary edema in the early stage, others were cured without sequelae.

CONCLUSIONSevere EV71 infection is more common in children younger than 3 years old, in which the profound complications include encephalitis and pulmonary edema. The mechanical ventilation should be critically urged for child with complicating cardiopulmonary failure as soon.

Child ; Child, Preschool ; Enterovirus A, Human ; classification ; Enterovirus Infections ; diagnosis ; therapy ; virology ; Female ; Hand, Foot and Mouth Disease ; diagnosis ; therapy ; virology ; Humans ; Infant ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo study the clinical features and treatment of serious brainstem encephalitis caused by enterovirus 71 (EV71) infection.

METHODSThe clinical data of 32 hospitalized children with serious brainstem encephalitis caused by EV71 infection between May and December 2008 were retrospectively reviewed.

RESULTSThe children whose age was younger than 3 years old accounted for 88% (22 cases). Fever(>38.5 degrees centigrade)lasting at least 3 days, frequent vomiting and limb twitch were presented as the main manifestations in the 32 children. Cyanosis, tachypnea, tachycardia and cold extremities were observed, and pulmonary edema or even pulmonary hemorrhage occurred in 8 children 3 to 4 days after the onset. The 32 children received a medical treatment: reduction of intracranial pressure with mannitol or frusemide, inhibition of inflammation reactivity with gamma globulin and methylprednisolone, and improvement of cardiac function and pulmonary edema with innotropic agents, fluid restriction and positive mechanical ventilation.

CONCLUSIONSVegetative nerve functional disturbance is the main clinical feature of brainstem encephalitis caused by EV71 infection in children. An early identification and treatment of pulmonary edema or hemorrhage is of great importance.

Brain Stem ; pathology ; Child, Preschool ; Encephalitis, Viral ; complications ; diagnosis ; therapy ; Enterovirus A, Human ; Enterovirus Infections ; complications ; diagnosis ; therapy ; Female ; Humans ; Infant ; Male ; Pulmonary Edema ; etiology ; therapy ; Retrospective Studies

BACKGROUND: Enterovirus is a common cause of aseptic meningitis, respiratory disease and nonspecific febrile illness. The conventional methods for laboratory diagnosis of enterovirus infections have been virus culture and serotyping by an immunofluorecent test. We studied a new and more rapid approach for enterovirus detection in cerebrospinal fluid (CSF) by real-time nested PCR. METHODS: This study was performed on 50 CSF specimens from patients suspected of aseptic meningitis. Enterovirus was detected in CSF by PCRs for 3 different targets and real-time nested PCR. Enterovirus culture was also performed in 44 CSF specimens. RESULTS: The positive rate of PCRs for each of the 3 different targets was 26.0%, 40.0%, or 46.0%, and that of real-time nested PCR was 86.0%. Only 6.8% were positive in culture. Thus, the positive rate of real-time nested PCR was much higher than other methods. CONCLUSIONS: Our study revealed that the real-time nested PCR should be useful for diagnosis of enterovirus infections because of a high sensitivity and rapid detection.
Cerebrospinal Fluid* ; Clinical Laboratory Techniques ; Diagnosis ; Enterovirus Infections ; Enterovirus* ; Humans ; Meningitis, Aseptic ; Polymerase Chain Reaction* ; Real-Time Polymerase Chain Reaction ; Reverse Transcription* ; Serotyping

Molecular detection of enterovirus (EV)71 RNA based on PCR methods is a quick and sensitive approach. At present, different PCR-based methods for EV71 RNA detection are available, but comparisons of results obtained using different approaches are limited. This study is to compare the analytical sensitivity and specificity of different real-time reverse transcription-polymerase chain reaction (rRT-PCR) and conventional reverse transcription-polymerase chain reaction (cRT-PCR) assays for enterovirus and EV71 detection, Altogether, three rRT-PCR assays and one cRT-PCR assay targeting the 5'UTR gene for universal detection of enterovirus; two rRT-PCR assays andone cRT-PCR assay targeting the VP1 gene for specific detection of EV 71 were examined. All assays showed good specificity. The detection sensitivity ranged from 8.19 x 10 to 8.19 x 10(5) copy equivalents. In general, rRT-PCR assays were more sensitive than cRT-PCR assays. All rRT-PCR assays showed 100% sensitivity for clinical specimens.
Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; virology ; Humans ; Real-Time Polymerase Chain Reaction ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo develop a method for detection of coxsackie B virus type 1-6 by RT-PCR.

METHODSA pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus.

RESULTSThis method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR.

CONCLUSIONThe RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.

DNA Primers ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin M ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods

BACKGROUNDTo establish a molecular detection and typing assay for identification and typing of human enteroviruses (HEV) which is suitable for clinical detection and epidemiologic research.

METHODSUsing both primers specific for HEV genus and HEV typing primers and reverse transcription polymerase chain reaction (RT-PCR) the authors detected preliminarily HEV by agarose gel electrophoresis and then identified serotype through nucleotide sequence analysis of RT-PCR amplicons. The monospecific antisera neutralization was applied to validate the typing results.

RESULTSThe serotype of 18 suspicious HEV samples was identified: 4 Coxsackievirus type A24 (CVA24), 3 CVB3, 1 CVB2, 1 CVA9, 1 CVA15, 1 Echovirus type 3 (E3), 1 E6, 1 E9, 1 E11, 1 E14, 1 E33 and 1 Rhinovirus type 9. The result was validated by monospecitive antisera neutralization.

CONCLUSIONThis RT-PCR assay for HEV detection and typing may be suitable for clinical detection and epidemic research since this method is sensitive and specific for direct identification and typing of HEV.

DNA Primers ; Enterovirus ; classification ; genetics ; Enterovirus Infections ; diagnosis ; Humans ; RNA, Viral ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Serotyping ; methods